Nucleosidesfi/Ides Abstracts

Abstract

Synthesis for Bridge-Oxygen Labeled Nucleoside Triphosphates.     

Population Differentiation in a Complex Bird Sound: A Comparison of Three Bioacoustical Analysis Procedures

We examined three bioacoustical analysis methods for comparing complex sounds among different populations. We chose the D‐syllable of the chick‐a‐dee call of the black‐capped chickadee (Poecile atricapilla) because it is a broadband sound representative of a class of vocalizations, common in many animals, that resists simple subjective classification for comparative studies. We examined the properties of the D‐syllable in field‐recorded samples from three different populations. The first method of data extraction sampled the amplitude values of a spectrum obtained in a single fast Fourier transform (SFFT) taken at the midpoint of each D‐syllable using multi‐speech software. The second method employed spectrogram cross‐correlation (SPCC) to obtain a matrix of similarity values between D‐syllables in the samples using canary software. The third method calculated similarity values obtained from the evaluation of four acoustic features of the D‐syllables derived from multi‐taper spectral analysis (MTSA) using sound analysis software. Following data extraction by these three techniques, we used multivariate statistical procedures to reduce the data for examination of differences among populations and to represent in scatter‐plots the patterns of clustering of the sounds. We found that the SFFT in the middle of the D‐syllable provided the poorest population discrimination following statistical processing, the SPCC method produced the next clearest population separation, and the MTSA method resulted in the most distinct separation of the three populations of D‐syllables. In carrying out these comparisons, we discovered that the characteristic environmental noise of a recording area can influence the signal properties of broadband sounds being compared by automated procedures, and could lead to faulty conclusions unless appropriate care is taken to mitigate the noise in which the signals of interest are embedded. Consequently we re‐analyzed our data following noise reduction and found less discrete population separation overall. However, the methods of SPCC and MTSA retained the ability to separate populations, with MTSA providing the sharpest discrimination among groups.     

Longitudinal Relationship between Elapsed Time in the Action Stages of Change and Weight Loss

Objective: The objective of this study was to examine the longitudinal relationship between the elapsed time in the action and maintenance stages of change for multiple target behaviors and weight loss or gain. Research Methods and Procedures: The research design was a prospective cohort study of overweight and obese primary care patients randomized to an obesity management intervention based on the Transtheoretical Model and a chronic disease paradigm. The target behaviors included increased planned exercise and usual physical activity, decreased dietary fat, increased fruit and vegetable consumption, and increased dietary portion control. The participants were 329 middle‐aged men and women with elevated body mass indices recruited from 15 primary care practices in Northeastern Ohio; 28% of the participants were African Americans. The main outcomes were weight loss (5% or more) or weight gain (5% or more) after 18 or 24 months of follow‐up. Results: There were significant (p < 0.05) longitudinal relationships between the number of periods (0 to 4) in action or maintenance for each of the five target behaviors, or a composite score taken across the five target behaviors, and weight loss. In all cases, there was a significant (p < 0.05) stepped (graded) relationship between the time in action or maintenance and weight loss (or gain). Discussion: The data support the concept of applying the Transtheoretical Model to the problem of managing obesity in primary care settings. The remaining challenge is to identify those factors that reliably move patients into the action and maintenance stages for long periods.     

Does signalling mitigate the cost of agonistic interactions? A test in a cricket that has lost its song

Prevailing models of animal communication assume that signalling during aggressive conflict mitigates the costs of fighting. We tested this assumption by staging dyadic encounters between male field crickets, Teleogryllus oceanicus, under three conditions: (i) both males could sing aggressive songs, (ii) neither male could sing, and (iii) one male could sing but the other could not. We conducted experiments on males from a Hawaiian population from Kauai that has recently evolved signal loss, and males from a Hawaiian population from the Big Island that has not. Among both populations, interactions between two silent males were characterized by higher levels of aggression than interactions involving one or two singing males. Because the level of aggression is strongly related to the cost of fighting, these data demonstrate that signalling mitigates the cost of fighting. In mixed trials, we found no statistically significant differences between the behaviour of calling and non-calling males in either population. We conclude that there is no evidence that the Kauai population exhibits special adaptations to alleviate the costs of signal loss. Finally, we found that males were much more likely to signal after their opponent''s retreat than after their own retreat. Aggressive song therefore meets the definition of a ‘victory display’.     

Evidence for the action of bovine follicular fluid factor(s) other than inhibin in suppressing follicular development and delaying oestrus in heifers.

The aim of this study was to investigate the importance of inhibin in the delay in return to oestrus in heifers induced by steroid-stripped bovine follicular fluid (bFF). Oestrous activity was synchronized in 18 Hereford x Friesian heifers with two injections of prostaglandin (PG) 12 days apart. At the time of the second PG injection (time 0), the animals were assigned at random to one of three experimental groups and received i.v. injections of 20 ml saline (controls, n = 6), whole bFF (FF group, n = 6) or bFF in which the bioactive inhibin content had been reduced by > 95% by immunoaffinity chromatography (-INH group, n = 6; inhibin content approximately 0.8 ml whole bFF) every 8 h for 2 days. In a dose-response study, 2.5 ml whole bFF was insufficient to delay oestrus consistently following a similar synchronization regimen. Blood samples were taken every 8 h, initially before each injection and then subsequently for a further 9 days for hormone analysis. Animals were observed every 8 h throughout the experiment for signs of behavioural oestrus. The ovaries of all animals were examined using real-time ultrasonography about 30 h after the second PG injection. Treatment failed to suppress peripheral follicle-stimulating hormone (FSH) concentrations, although a significant increase was observed in both treatment groups after cessation of injections. Progesterone concentrations fell immediately after the second PG injection in all animals and remained below minimum detectable concentrations in all treated animals for the remainder of the experiment. In control animals, progesterone rose above minimum detectable concentrations by day 6 and continued to rise until the end of the experiment. Analysis of samples taken from treated animals several days after observed oestrus revealed that all had apparently ovulated. Mean daily luteinizing hormone (LH) concentrations did not differ between treatment groups before ovulation, but after ovulation, mean daily LH was significantly reduced in control animals as progesterone concentrations rose. Follicular development, as assessed by the mean antral diameter of the largest follicle on a pair of ovaries at ultrasound examination, was significantly suppressed in treated animals compared with controls (P < 0.01) and there was no significant difference (P = 0.397) between the two treatment groups. Control animals displayed oestrus 68 h (+/- 8 SEM) after the second PG injection, but oestrus was delayed in treated animals to 186h +/- 5 (FF group) and 191 h +/- 6 (-INH group).     

The release of bradykinin in bovine mastitis.

The kinin peptides are released during inflammation and are amongst the most potent known mediators of vasodilatation, pain and oedema. Despite early reports of the presence of kinins in milk, no previous study has investigated the role of the kinin system in bovine mastitis. The present study indicated that mastitis was accompanied by raised levels of bradykinin (BK) in milk and the increased levels of BK correlated with the severity of mastitis. Raised BK levels in mastitic milk were not dependent on the presence of inflammatory cells, nor were they secondary to changes in blood levels of BK. In milk from sub-clinically inflamed quarters, BK was raised in those milks where Staphylococcus aureus (S. aureus) was isolated but not in those milks where no pathogen was isolated. Increasing S. aureus artificially, also caused an increase in the milk BK. Increases in milk BK were not restricted only to the mastitic quarters of the udder. In udders in which mastitis was detected in one or more quarters, BK increases were also detected in the apparently uninvolved quarters.     

Subcellular localization of 3-methylcrotonyl-coenzyme A carboxylase in bovine kidney

The intracellular localization of 3-methylcrotonyl-CoA carboxylase (MCase), an enzyme of the leucine oxidative pathway, was studied in bovine kidney. Differential centrifugation of kidney homogenates demonstrated that the majority of the enzyme was associated with the mitochondrial and cytosolic fractions. Isopycnic centrifugation of the mitochondrial fraction demonstrated cofractionation of MCase with mitochondrial markers, but not with lysosomal markers, consistent with a mitochondrial location for the enzyme. Using different homogenization techniques and comparing the fractional extraction of MCase and mitochondrial and cytosolic marker enzymes, the appearance of MCase in the “cytosolic” fraction was shown to be due to mitochondrial damage. The intramitochondrial distribution of MCase was determined using a digitonin procedure, and indicated that the enzyme is associated with the inner mitochondrial membrane. Although a fraction of MCase (30–40%) was “solubilized” by homogenization of whole mitochondria, the remaining MCase (60–70%) was tightly associated with the mitochondrial membrane fraction. Release and “solubilization” of this latter fraction was achieved by polyethylene glycol treatment. The “solubilized” MCase was stabilized in a glycerol-containing buffer.     

Molecular cloning and functional characterization of KCC3, a new K-Cl cotransporter

We isolated and characterized a novelK-Cl cotransporter, KCC3, from human placenta. The deduced proteincontains 1,150 amino acids. KCC3 shares 75-76% identity at theamino acid level with human, pig, rat, and rabbit KCC1 and 67%identity with rat KCC2. KCC3 is 40 and 33% identical to twoCaenorhabditis elegans K-Cl cotransporters and ~20%identical to other members of the cation-chloride cotransporter family(CCC), two Na-K-Cl cotransporters (NKCC1, NKCC2), and the Na-Clcotransporter (NCC). Hydropathy analysis indicates a typical KCCtopology with 12 transmembrane domains, a large extracellular loopbetween transmembrane domains 5 and 6 (unique to KCCs), and largeNH₂ and COOH termini. KCC3 is predominantly expressed inkidney, heart, and brain, and is also expressed in skeletal muscle,placenta, lung, liver, and pancreas. KCC3 was localized to chromosome15. KCC3 transiently expressed in human embryonic kidney (HEK)-293cells fulfilled three criteria for increased expression of K-Clcotransport: stimulation of cotransport by swelling, treatment withN-ethylmaleimide, or treatment with staurosporine.